Tissue-holder receptacles and method of preparing tissue for microscopic examination



Feb. 26, 1957 E. c. WHITEHEAD 2,783,180

TISSUE-HOLDER RECEPTACLES AND METHOD 0F PREPARING TISSUE FOR MICROSCOPICEXAMINATION Filed April 14, 1954 FIG.: f4

FIG. 3

A" INVENToR.

\ ATTORNEYS Edwin C. Whitehead, Crestwood, N. Y., assigner to TechniconInternational, Ltd., Bronx, N. Y., a corporation of New York ApplicationApril 14, 1954, Serial No. 423,186

23 Claims. (Cl. 167-845) This invention relates to the preparation of.human or animal tissue for microscopic examination and more particularlyto `containers or holders which are employed for holding the tissuespecimens during their immersion in the various liquids, i. e.,tixatives, dehydrants, inltrat ing agents, clearing agents, etc., usedin the preparation of the tissue.

Tissue processing apparatus of the type which may be used to advantagefor immersing tissue in the various liquids is shown, for example, inPatents Nos. 2,341,197 and 2,341,198, granted to Edwin C. Weiskopf,United States Letters Patent No. 2,583,379, granted to Nelson G. Kling,and in United States Patent No. 2,741,221, granted to Edwin C..Weiskopfand Andres Ferrari, Jr. The aforesaid patents and patent application areowned by the assignee of the present application. In the operation ofsuch apparatus the tissue holders are moved automatically into and outof each of a plru'ality of liquidcontaining receptacles, in succession,with intervening intervals during which said holders are positionedwithin the receptacles respectively for subjecting the tissue or othermaterial to the action of the various liquids provided in saidreceptacles, i. e. lixatives, dehydrants, clearing agents, infiltratingagents, etc. Y v

One object of the invention is to prevent the curling or otherdistortion of tissue, for example intestines or other tubes or any othertissue Ispecimens of thin wall section which is likely to curl up orbecome distorted by the fixative or other tissue processing liquids. Inthis connection it will be understood that if the tissue is allowed tocurl or become distorted, it is diilicult or impossible to cut a truerepresentative section from the paraffin block in which the tissue isembedded. The object just stated is to eliminate this serious difficultyand disadvantage. q Y

Another Vobject isV the provision of a tissue holder receptacle whichenables the tissue specimen to be disposed in optimum condition fortreatment by the processing liquids.

A further object is the provision` of a tissue-holder receptacle whichfacilitates the handling of the tissue by the pathologist or thetechnician in the laboratory.

A yet further object is generally to provide a tissueholder receptaclewhich is inexpensive and at the same time superior in many` respects toknown tissue-holder receptacles.

The above objects of the invention and other objects ancillary theretowill be fully understood from the following description considered inconnection with the accompanying drawings, which are to be considered asillustrative of the invention but not in limitation thereof.

In the drawings: i l,

Fig. 1 is a vertical sectional view, partly in elevation,illustratingpart'of an automatic immersion apparatus for practising thepresent invention;

Fig. 2 is a horizontal sectional view, on a larger scale,

various Sevresr Patent Y, matically adheres tothe surfacethereof, the`adhesion'of 2,783,180 Patented Feb. 26, 1957 ICC taken on line 2 2 ofFig. l, the top part of one of the material holders being cut away forillustrative purposes; Fig. 3r is a vertical sectional View taken on theline 3--3 of Fig. 2; Y

Fig. 4 is a perspective view of one of the material holders, its coverbeing removed; Y

Fig. 5 is a perspective view of the cover for the rnaterial holder; and,

Fig. 6 is a side View of two tissue holders, illustrating a modication.Y

Referring first to Fig. l, a plurality of material holders 10 are shownimmersed in a beaker or other liquid receptacle 12, which is carried bythe novel carrier 14 which is more specifically described and claimed inthe co-pending application of Edwin C. Weiskopf and Andrs Ferrari, lr.,Serial No. 423,187, tiled concurrently herewith, and owned by theassignee of this application.Y

Carrier 14 and receptacles 12 are well adapted for use with automaticimmersion apparatus 16, of the type shown in the aforesaid co-pendingapplication of Nelson G. Kling and by the above mentioned patents.

Pursuant to the present invention provision is made in the tissue-holderreceptacle for automatically holding the tissue specimen in flatcondition so that it will not curl up or become distorted, and thisresult is accomplished by making the cover, in whole or in part, of thereceptacle of a material to which the tissue specimen will self-adhereso that the tissue will stay flat While it is processed in thevariousliquid agents.

Referring now to Figs. 2 to 5 in detail, which illustrates the presentlypreferred mode of practicing the invention, the body portion of thetissue holder receptacle is preferably non-metallic and can be formed,for example and not by Way of limitation, of a plastic or ofpapier-mache, pressed paper, etc., and is generally cupshaped having abottom 20, provided with a plurality of apertures 22 to provide accessof the liquid to the tissue when the receptacles are immersed in theliquids in the beakers 12, and a peripheral side wall 24. Said side wall24 includes a lower or main portion 26 and an integral upper portion 28which is outwardly ofset to provide a shoulder Sil as best illustratedin Fig.f3. The upper portion 28 is provided with an internal peripheralgroove 32 adjacent shoulder 311 for inserting the peripheral marginaledge of a cover 34. j

In accordance with this invention, the tissue is mounted on the innerside of the cover 34, as indicated at T in Fig. 3, Awhich shows a pieceof the tissue mounted on the cover. In order to hold the tissue specimenin flat condition and prevent its curling up, or becoming distorted whenvtreated bythe xative or other tissue processing liquids, said cover hasan inner surface layer which is of such nature that the tissue willadhere to it, without the use of an extraneous adhesive, when placed incontact therewith. For this purpose the cover has an inner layer 46formed of porous or absorptive material, such as for example theblotting surface layer of an ink .blotteu The outer layer 48 of cover 34may be relatively stili.C and isl upper surface of a conventional inkblotter. Layers 46' and 48- are laminated and joined to each other in4surfaceto-surface relation by a glue which is preferably insoluble inor resistant to water ror in the agents in which the tissue isprocessed, such as for example a phenolic glue. The piece of tissue 49is laid flat on portion 46 and autothe tissue 49 to portion 46 beingaided by the albumin normally present in the tissue.' Perforations 56may be' provided in cover 34 in orderto augment access of the tissueprocessing liquids to the tissue. However, these may be omitted from thecover. Also it desired wall 24 may be perforated.

In use, the tissue specimen in itsV moist or in a moistened condition isplacedon layer 46 and will adhere thereto. The adhesion is enhanced bythe action of the xative on the albumin inherently present in thetissue, as a result of which the albumin is coagulated by theformaldehyde iixative and there is a releasable bond between the tissueand the fibres of the porous material of layer 46. The tixativemay beapplied to the tissue specimen, for this lpurpose, as soon as the tissueis applied to the cover, or after the'cover is'placed on the bodyportion of the receptacle 10. Thereafter a plurality of the receptacleswith similar cover-carrying tissue are mounted on the carrier.14,.asdescribed in the application tiled concurrently herewith, or may beplaced in a perforated basket, as described hereafter, and in the courseof the operation of the apparatus the tissue will be acted upon by thevarious liquids in the several beakers. The first liquid is the iixativeand this will ordinarily be suiiicient to improve the securement of thetissue to the cover without a preliminary treatment of the-tissue withthe iixative when the tissue isrst placed on the cover.

As illustrated in Fig. 6, the tissue-holder receptacles may have theirlower peripheral edges corrugated as indicated at 52, for example, toprevent any possibility of the liquids in which the holders are immersedfrom not passing freely into and out of they holders through the bottomthereof when the holders are mounted one on top of the other asillustrated in Figs. 1 to 3. Ordinarily however, the failure of anadequate quantity of liquid to circulate freely into and out of theholders through the bottom openings is remote or negligible, especiallyso when the holders are used in processing machines of the type shown inthe above mentioned Patent No. 2,741,221 according to which the tissueholders are vertically reciprocated. The tissueholder receptacles shownin Fig. 6 and designated 10A are the same in all respects as holders 10described above.

It will be understood that when the tissue 49 is applied tothe surface46 of cover 34, and then immersed in each of the liquids including theparain baths, tissue specimens, such as pieces of intestines, tubes orany thin wall sections, or anything which is likely to curl up or becomedistorted after treatment by the fixative or the other tissue processingliquids, will hereafter when processed with the novel material holder 10or 10A and cover 34 in accordance with the above technique as set forthin accordance with the present invention produce a substantially flatnon-curled specimen. The tissue specimen produced in accordance with thepresent invention will be imbedded in the parafin block in. asubstantially flat condition whereby truly representative sections ofthe tissue specimen can then be cut from said paraffin block, afterwhich the tissuespecimens can be deparatlinized, stained and mounted onslides.

As clearly illustrated in Fig. 4, the upper peripheral edge of the upperportion 28 of wall 24 has an ear 0r extension member 36 having a portion38 provided with an aperture as at 40. Member 36 is provided with ascore line at 42, whereby the portion 38 may be severed from portion 44of flange member 36, the purpose of which will be hereinafter furtherexplained.

It will be understood that the tissue processing machines have aplurality of receptacles arranged in a circular row on one or morehorizontal supports, indicated herein at 62 and 64, and that the presentdrawings show only a part of a machine of the type shown by the abovementioned Kling patent and the Weiskopf and Ferrari Patent No.2,741,221, further illustration or description beingunnecessarytherein..y However, `itmay benotedA that in order to impartthe various movements for processing the tissue specimens, such as theraising and lowering of the material holders'10 from the receptacles 12containing the tissue processing liquids, and for imparting the verticalreciprocating movement to material holders 10, a conveyor 68 isprovided. Said conveyor 68 includes the member or spider which ispartially illustrated at 70, rods 71 and 72 suitably secured to spider79 in any suitable manner. At the outer end thereof, each of the rods 71and 72 is provided with a bracket or arm 74, 74 one end of which isprovided with a sleeve 75 for securement to the free end of said rods.The cnrrier 14 is supported by a spindle 76 which has suitable provisionfor mounting on `the other end of arm 74. Spindle 76 is mounted by meansof member 78 which has an enlarged head. Mounted on the other end ofspindle 76 is a channel shaped member 80 for carrying U-shaped member 82of carrier 14, the leg portions of which are provided with pinprojections 84, 84. The carrier 14 which is disclosed in the co-pendingapplication of Edwin C. Weiskopf and Andres Ferrari, Ir., Serial No.423,187, tiled concurrently herewith, which is shown immersed in upperreceptacle 12 also includes the supporting spindle 86 which is connectedto cross-bar SS, said cross-bar being apertured at either end forreceiving the pin projections 84. Also mounted on spindle 76 is a cover9i) for receptacles 12. As set out in the previously noted co-pendingapplication which is tiled concurrently herewith, said supportingspindle 86 comprises at least two portions in one modification, whichcan be readily disconnected from each other whereby to place materialholders 10'or 10A thereon, after which said portions can again beconnected whereby said material holders 1d or 10A when so placed on saidsupporting spindle 86 can be readily reciprocated within the liquids insaid receptacles 12 and can also be readily moved from one receptacle toanother.

As noted in Figs. 1 and 2, a plurality of the material holders 10 or 10Amay be mounted on spindle 86, said material holders being mounted onsaid spindle by means of ear 36, and as noted previously, the lowerportion of spindle 86 can be detached from the upper portion whereby thelower portion of spindle 86 can then be inserted through aperture 40,and then again attached to `the upper portion of spindle 86, whereby theplurality of material holders 10 or 10A can be readily coupled to thecarrier 14 for imparting to material holders 10 or 10A, thereciprocating movement required for the processing of the tissuespecimen.

Pursuant to another feature of the present invention, material holders10 or 10A may also be used with the basket type holder 92 by simplyplacing material holders 10 or 10A therein. As illustrated in Fig. l,said basket type holder 92 is disposed within the lower receptacle 12,which is supported on the lower base 64.` As` illustrated in theaforesaid patents and as disclosed herein, said basket type holder 92shown on the lower shelf 64, for illustrative purposes, is connected toand supported by the U-shaped member 82 by pins 84. As noted pre`viously, ange 36 is provided with a score line at 42 whereby to severportion 38 from 44. When material holders 10 or 10A are used with thebasket type holder 92, there is no need for the portion 38, andtherefore, portion 38 may be removed if desired.

in the aforesaid copending application tiled concurrently herewith,Serial No. 423,187, a material holder carrer mechanism is disclosed andclaimed, wherein tracks are provided on the carrier 14 by modifying thespindle portion 86. Flange 36 of material holders 1d or 10A may also bemodified in accordance with the aforesaid copending application so as tohave provided a complementary portion for engagement with the tracks.

As noted in Figs. 1 and 3,V the material holders 1t) or 10A are shown instacked relation; however, as a result of thereciprocatingunovernentimparted to carrier 14 or the basket type carrier 92 by the immersionapparatus 16, said materialholders will ordinarily have` relativemovement imparted thereto, as previously explained. Portion 38 has anaperture 40 which is of a slightly larger diameter than the diameter ofsupporting spindle 86 whereby to allow the material holders 10 or 10A tomove relative to each other. The tissue processing liquids can fiowthrough the apertured base through apertures 22 so as to infiltratetissue specimen.

While I have shown and described the preferred embodiment of myinvention, it will be understood that various changes may be madetherein without departing from the underlying idea or principles of theinvention within the scope of the appended claims.

Having thus described my invention, what I claim and desire to secure byLetters Patent, is:

v1. A tissue-holder receptacle for use in treating the tissue with aprocessing liquid in the preparation of the tissue for microscopicexamination, comprising a container having an internal surface partthereof formed of porous fibrous material for mounting the tissuethereon in position for access thereto of processing liquid within thecontainer, said porous fibrous material having a plurality of openingstherethrough for access of said liquid to the tissue.

2. A tissue-holder receptacle for use in treating the tissue with aprocessing liquid in the preparation of the tissue for microscopicexamination, comprising a container having a wall provided with aplurality of openings therethrough for the pasasge of said liquid intoand out of the container, said container having an internal surface partthereof formed of porous fibrous material for mounting the tissuethereon in position for access thereto of processing liquid enteringsaid container through said openings, said container having a partconfronting said internalsurface part in spaced relation thereto anddefining therewith a space for the tissue mounted on said porous fibrouspart. A

3. A tissue-holder receptacle for use in treating the tissue with aprocessingliquid in the preparation of the tissue for microscopicexamination, comprising a container having a internal surface partthereof formed of porous fibrous material for mounting the tissuethereon in position for access thereto of processing liquid within thecontainer, said container having a removable cover provided with saidinternal fibrous surface.

4. A tissue-holder receptacle for use in treating the tissue with aprocessing liquid in the prepaartion of the tissue for microscopicexamination, comprising a container having an internal surface partthereof formed of porous fibrous material for mounting the tissuethereon in position for access thereto of processing liquid within thecontainer, said container having a removable cover provided with saidinternal fibrous surface, said cover having a moisture-resistant outersurface.

5. A tissue-holder receptacle for use in treating the tissue with aprocessing liquid in the preparation of the tissue for microscopicexamination, comprising a perforated container having an internalsurface part thereof formed of porous fibrous material for mounting thetissue thereon in position for access thereto of processing liquid,entering said container through the perforations provided therein, saidcontainer having a removable cover provided with said internal fibroussurface.

6. A tissue-holder receptacle for use in treating the tissue with aprocessing liquid in the preparation of the tissue for microscopicexamination, comprising a perforated container having an internalsurface part thereof formed of porous fibrous material for mounting thetissue thereon in position for access thereto of processing liquid,entering said container through the perforations provided therein, saidcontainer having a removable cover provided with said internal fibroussurface, said cover hav` ing a moisture-resistant outer surface.

7.1 A tissue-holder receptacle for use in treating the tissue with aprocessing liquid in the preparation of the tissue for microscopicexamination, comprising a container having a internal surface partthereof formed of porous fibrous material for mounting the tissuethereon in position for -access thereto of processing liquid within thecontainer, said container having a removable cover provided with saidinternal fibrous surface, said container having a wall thereof providedwith perforations for the passage of liquid into andout of thecontainer.

8. A tissue-holder receptacle for use in treating the tissue with aprocessing liquid in the preparation of the tissue for microscopicexamination, comprising a container having an internal surface partthereof formed of porous fibrous material for mounting the tissuethereon in position for access thereto of processing liquid within thecontainer, said container having a removable cover provided with saidinternal fibrous surface, said container having a bottom Wall providedwith perforations for the passage of liquid into and out ofthecontainer, said cover having an inner layer of porous paper providingsaid in v ternal porous surface part of the container, an outer layerjoined to said porous layer, and a layer of water insoluble adhesiveinterposed between said inner and outer layers securing them to eachother and preventing the passage of moisture from said inner layer tosaid outer layer.

9. A tissue-holder receptacle for use in treating the tissue with aprocessing liquid in the preparation of the tissue for microscopicexamination, comprising a container having an internal surface partthereof formed of porous fibrous material for mounting the tissuethereon in position for access thereto of processing liquid within thecontainer, said container having a removable cover provided with saidinternal fibrous surface, said cover having an inner layer of porouspaper providing said internal porous surface part of the container, anouter layer joined to said porous layer, and a layerof water insolubleadhesive interposed between said inner and outer layers securing them toeach other and preventing the passage of moisture from said inner layerto sa-id outer layer.

10. A tissue-holder receptacle for use in treating the tissue with aprocessing liquid in the preparation of the tissue for microscopicexamination, comprising a container having aV body portion provided withperforations, theupper part of said body portion having an internalcover-seat, and a` cover for the top of container releasably secured onsaid internal cover-seat of said body portion, said cover having aninternal fibrous surface for carrying the tissue thereon.

ll. A tissue-holder receptacle for use in treating the tissue with aprocessing liquid in the preparation of the tissue for microscopicexamination, comprising a container having a body portion provided withperforations, the upper part of said body portion having an internalcover-seat, and a cover for the top of container releasably secured onsaid internal cover-seat of said body portion, said cover having aninternal porous fibrous surface for mounting the tissue thereon inposition for access thereto of processing liquid within the container.

` portion, said cover having an internal porous fibrous sur face formounting the tissue thereon in position for access thereto of processingliquid within the container, and a lateral extension on said bodyportion externally thereof having a formation for mounting the tissueholder in a tissue processing machine.

13. The method of preparing tissue for microscopic examinationzaccordingto which the tissue is immersed in a processing liquid, comprisingmounting the tissue specimen in at condition on a member having a porousbrous surface to which the tissue adheres and immersing said mountedtissue specimen in the processing liquid.

14. The method of preparing tissue for microscopic examination accordingto which the tissue is immersed in a processing liquid, comprisingmounting the tissue speci men in fiat condition on a member having aporous fibrous surface to which the tissue adheres and immersing saidmounted tissue specimen in a liquid iixative for coagulating the albuminin the tissue and thereby increasing the adhesion of the tissue to saidsurface.

15. The method of prepaling tissue for microscopic examinationaccordingto which the tissue is immersed in a processing liquid,comprising mounting the tissue specimen in at condition on a memberhaving a porous fibrous surface, and applying a coagulant to the tissueon said member to form a releasable adhesion of the tissue to saidsurface.

16. A tissue-carrier for use in treating the tissue with a processingliquid in the preparation of the tissue for microscopic examination,comprising a member having a porous surface to mount the tissue specimenin selfadhering relation thereon and having provision for the passage ofliquid therethrough for exposing the tissue to the processing liquid foraction of said liquid thereon.

17. A tissue-carrier for use in treating the tissue with a processingliquid in the preparation of the tissue for microscopic examination,comprising a member having a porous surface to mount the tissue specimenin selfadhering relation thereon and exposed to the processing liquidfor action of said liquid thereon, and a receptacle member adapted toenclose the tissue and provided with openings for the passage of saidliquid to the tissue on said first mentioned member.

18. A tissue-carrier for use in treating the tissue with a processingliquid in the preparation of the tissue for microscopic examination,comprising a member having a porous surface to mount the tissue specimenin selfadhering relation thereon and exposed to the processing liquidfor action of said liquid thereon, and a receptacle member adapted toenclose the tissue and provided with openings for the passage of saidliquid to the tissue on said first mentioned member, one of said membershaving a lateral extension provided with means for mounting the tissuecarrier on a support.

19. A tissue-carrier for use in treating the tissue with a` processingliquidin the preparation of the tissue for microscopic examination,comprising a member formed of sheet material and having a porous surfaceto mount the tissue specimen in self-adhering relation thereon, saidmember having provision for exposing opposite surfaces ofthe tissue tothe processing liquid.

20. A removable cover for a tissue-holder receptacle for use with aprocessing liquid in the preparation of the tissue for microscopicexamination, said cover comprising a member formed of sheet material andhaving an internai surface part formed of fibrous material for mountingthe tissue thereon, and said cover being perforated to provide foraccess of the processing liquid to said tissue.

21. A removable cover for a tissue-holder receptacle for use with aprocessing liquid in thc preparation of the tissue for microscopicexamination, said cover comprising a member formed of sheet material andhaving its internal surface formed of brous material for mounting thetissue thereon and its external surface formed of relatively moistureresistant material, and said cover being perforated to provide foraccess of the processing liquid to said tissue.

22. A tissue-holder receptacle for use in treating the tissue with aprocessing liquid in the preparation of the tissue for microscopicexamination, comprising a body portion and a cover therefor, saidreceptacle body and cover providing a tissue enclosing and retainingchamber, and having an inner porous surface portion for attaching thetissue thereto within said chamber, said receptacle having provision forthe passage of liquid into and out of said chamber for access of theprocessing liquid to the tissue therein.

23. A tissue-carrier for use in treating the tissue with a processingliquid in the preparation of the tissue for microscopic examination,comprising a paper disk having a porous surface to mount the tissuespecimen in selfadhering relation thereon, said disk having perforationstherethrough to provide access of the processing liquid to the adheredsurface of the tissue.

References Cited in the le of this patent UNITED STATES PATENTS1,236,832 Frye Aug. 14, 1917 2,393,580 Weiskopf Mar. 31, 1942 2,539,802Weiskopf Jan. 30, 1951 2,601,307 Meyer et al. June 24, 1952

13. THE METHOD OF PREPARING TISSUE FOR MICROSCOPIC EXAMINATION ACCORDINGTO WHICH THE TISSUE IS IMMERSED IN A PROCESSING LIQUID, COMPRISINGMOUNTING THE TISSUE SPECIMEN IN FLAT CONDITION ON A MEMBER HAVING APOROUS FIBROUS SURFACE TO WHICH THE TISSUE ADHERES AND IMMERSING SAIDMOUNTED TISSUE SPECIMEN IN THE PROCESSING LIQUID.